CAJ | 학술논문

目的以T淋巴细胞在甲状腺相关性眼病(TAO)发生发展中的重要作用作为切人点,筛选出具有意义的microRNA.方法实验研究.收集对照组、TAO静止期组和TAO急性期组各3例患者的外周静脉血,提取人外周血单个核细胞(PBMC),采用新一代高通量测序(NGS),筛选出TAO患者(包括TAO静止期组和TAO急性期组)与对照者差异表达的microRNA.然后采用实时荧光定量PCR技术,在对照组、TAO静止期组和TAO急性期组各5例的扩大样本中对上述筛选出的microRNA予以t检验验证.结果根据测序结果筛选出差异有统计学意义,且相差倍数＞2的microRNA,TAO静止期组与对照组比较共18个,TAO急性期组与对照组比较共31个.2个TAO患病组共同与对照组表达差异有统计学意义的microRNA为miR-19a-5p、miR-30c-2-3p、miR-548c-5p、miR-6718-5p,且均呈现明显上调趋势.实时荧光定量PCR验证结果与测序结果基本相符.结论 TAO患者PBMC中存在microRNA表达差异,miR-19a-5p、miR-30c-2-3p、miR-548c-5p、miR-6718-5p是潜在的TAO免疫诊断学标记物,部分可能参与TAO的免疫学发生发展机制.
목적 이T림파세포재갑상선상관성안병(TAO)발생발전중적중요작용작위절인점,사선출구유의의적microRNA.방법 실험연구.수집대조조、TAO정지기조화TAO급성기조각3례환자적외주정맥혈,제취인외주혈단개핵세포(PBMC),채용신일대고통량측서(NGS),사선출TAO환자(포괄TAO정지기조화TAO급성기조)여대조자차이표체적microRNA.연후채용실시형광정량PCR기술,재대조조、TAO정지기조화TAO급성기조각5례적확대양본중대상술사선출적microRNA여이t검험험증.결과 근거측서결과사선출차이유통계학의의,차상차배수＞2적microRNA,TAO정지기조여대조조비교공18개,TAO급성기조여대조조비교공31개.2개TAO환병조공동여대조조표체차이유통계학의의적microRNA위miR-19a-5p、miR-30c-2-3p、miR-548c-5p、miR-6718-5p,차균정현명현상조추세.실시형광정량PCR험증결과여측서결과기본상부.결론 TAO환자PBMC중존재microRNA표체차이,miR-19a-5p、miR-30c-2-3p、miR-548c-5p、miR-6718-5p시잠재적TAO면역진단학표기물,부분가능삼여TAO적면역학발생발전궤제.
Objective To compare the primary screen differential expression of microRNAs in thyroid-associated ophthalmopathy (TAO) groups and a control group with partial immunopathogenesis.Methods Experimental study.Patients were divided into 3 groups:a silent TAO group (3 samples),an acute TAO group (3 samples) and a control group (3 samples).After all microRNAs were extracted from the peripheral blood mononuclear cells (PBMC) of each sample,the next generation sequence (NGS) was examined and used to establish the microRNA profile.The differential expression of microRNA was found by comparing the silent TAO group vs.the control group and the acute TAO group vs.the control group.MicroRNAs were expressed differently in the groups.Then quantitative real-time PCR (qrt-PCR) was used to verify those differences in another 5 samples from each group.Results Compared to the control group,18 microRNAs in the silent TAO group and 31 in the acute TAO group were sorted out by setting a criterion of P＜0.05 and a fold change (fc)＞2.There were 4 microRNAs (miR-19a-5p,miR-30c-2-3p,miR-548c-5p,miR-6718-5p) in common between the two comparisons.The up-regulation trends of those 4 microRNAs verified by qrt-PCR were in line with the results of the microRNA-seq.Conclusion Differential expressed microRNAs in PBMC could be novel biomarkers for detection of TAO in humans and some may play a key role in the immunopathogenesis of TAO.